Androgen receptor gene status in plasma DNA associates with worse outcome on enzalutamide or abiraterone for castration-resistant prostate cancer: a multi-institution correlative biomarker study

2632A>G (p.T878A)] were observed in eight (11%) post-docetaxel but no chemotherapy-naı¨ve abiraterone-treated patients and were also associated with worse OS (HR 3.26; 95% CI 1.47-not reached; P¼0.004). There was no interaction between AR and docetaxel status (P¼0.83 for OS, P¼0.99 for PFS). In the PREMIERE trial, 11 patients (12%) with AR gain had worse PSA-PFS (sPFS) (HR 4.33; 95% CI 1.94-9.68; PA (p.L702H) and 520 3

Androgen receptor gene status in plasma DNA associates with worse outcome on enzalutamide or abiraterone for castration-resistant prostate cancer: a multi-institution correlative biomarker study.

Background There is an urgent need to identify biomarkers to guide personalized therapy in castration-resistant prostate cancer (CRPC). We aimed to clinically qualify androgen receptor (AR) gene status measurement in plasma DNA using multiplex droplet digital PCR (ddPCR) in pre- and post-chemotherapy CRPC.Methods We optimized ddPCR assays for AR copy number and mutations and retrospectively analyzed plasma DNA from patients recruited to one of the three biomarker protocols with prospectively collected clinical data. We evaluated associations between plasma AR and overall survival (OS) and progression-free survival (PFS) in 73 chemotherapy-naïve and 98 post-docetaxel CRPC patients treated with enzalutamide or abiraterone (Primary cohort) and 94 chemotherapy-naïve patients treated with enzalutamide (Secondary cohort; PREMIERE trial).Results In the primary cohort, AR gain was observed in 10 (14%) chemotherapy-naïve and 33 (34%) post-docetaxel patients and associated with worse OS [hazard ratio (HR), 3.98; 95% CI 1.74-9.10; P A (p.L702H) and 2632A>G (p.T878A)] were observed in eight (11%) post-docetaxel but no chemotherapy-naïve abiraterone-treated patients and were also associated with worse OS (HR 3.26; 95% CI 1.47-not reached; P = 0.004). There was no interaction between AR and docetaxel status (P = 0.83 for OS, P = 0.99 for PFS). In the PREMIERE trial, 11 patients (12%) with AR gain had worse PSA-PFS (sPFS) (HR 4.33; 95% CI 1.94-9.68; P < 0.001), radiographic-PFS (rPFS) (HR 8.06; 95% CI 3.26-19.93; P < 0.001) and OS (HR 11.08; 95% CI 2.16-56.95; P = 0.004). Plasma AR was an independent predictor of outcome on multivariable analyses in both cohorts.Conclusion Plasma AR status assessment using ddPCR identifies CRPC with worse outcome to enzalutamide or abiraterone. Prospective evaluation of treatment decisions based on plasma AR is now required.Clinical trial number NCT02288936 (PREMIERE trial)

Estudios de Caso sobre Ciencias Agropecuarias y Rurales en el siglo XXI.

Libro científico sobre estudios de casos en el medio agropecuario y ruralCon el advenimiento del siglo XXI y el avance de los procesos de globalización, el medio rural presenta diversos cambios económicos, sociales, políticos y culturales. Lo anterior significa que el campo es un objeto de estudio altamente dinámico, complejo e inasible. las ciencias agropecuarias y rurales, en la actualidad, requieren de un abordaje sistémico e interdisciplinario que den cuenta de la heterogeneidad de situaciones y contextos que enfrenta el campo mexicano. La presente obra agrupa 18 estudios de caso, que capturan algunas fotografías de las diversas problemáticas de la ruralidad mexicana, con lo cual se pretende dar cuenta tanto de los objetivos de estudio como de la perspectiva teórico metodológico desde que estos son abordados. lo anterior tiene que ver con el hecho de que las ciencias agropecuarias y rurales manifiestan un alto grado de observación empírica, motivo por el que los estudios de caso se convierten en la perspectiva metodológica idónea que permite ir y venir de la realidad a la teoría y viceversa para la construcción de objetos de estudio. En este volumen se aborda una gran diversidad de casos, que sintetizan la heterogeneidad de enfoques y perspectivas mediante las cuales los fenómenos agropecuarios y rurales han sido abordados en el Instituto de Ciencias Agropecuarias y Rurales de la Universidad Autónoma del Estado de México, en los últimos 30 años

A la croisée de la division cellulaire et de la différenciation cellulaire : rôle de Seh1 dans les cellules souches embryonnaires de souris

Les complexes des pores nucléaires (NPCs) sont des larges structures macromoléculaires ancrées dans l'enveloppe nucléaire, et composées d'environ 30 protéines différentes appelées nucléoporines ou Nups. Au-delà de leur rôle critique dans le transport des molécules, les NPCs participent à une grande variété de processus biologiques, tels que la division cellulaire, l'organisation de la chromatine et la régulation de l'expression des gènes. Le principal sous-complexe structural des NPCs est le complexe Y-complexe, qui est composé de 9 nucléoporines distinctes. Le Y-complexe est aussi localisé aux kinétochores pendant la mitose où il régule la congression et la ségrégation des chromosomes. Une nucléoporine de ce complexe, Nup133, a été impliquée dans le développement embryonnaire de la souris. Nup133 n’est pas requise pour la survie des cellules souches embryonnaires murines (mESCs) à l'état pluripotent mais devient essentielle lors de leur différenciation. Le Y-complexe joue donc un rôle majeur dans de multiples processus biologiques mais le rôle spécifique de chacune de ses sous-unités n’a pas été étudié en profondeur. En particulier, une nucléoporine, Seh1, est critique pour la localisation du Y-complexe et de plusieurs régulateurs mitotiques clés, notamment Aurora B aux kinétochores. Dans ce contexte, Seh1 apparaît comme un candidat intéressant, non seulement en raison de son implication en mitose, mais aussi parce qu'elle appartient à un second sous-complexe, nommé GATOR2, qui régule indirectement la voie mTORC1. On ne sait toutefois pas si les fonctions mitotiques de Seh1 reposent sur sa localisation au niveau du pore, des kinétochores ou du complexe GATOR2. Les travaux réalisés au cours de cette thèse ont révélé que les mESCs Seh1-/- sont viables à l'état indifférencié, mais que leur viabilité est gravement compromise lors de la différenciation. Nous avons constaté que l'inactivation de Seh1 est associée à un retard dans la progression mitotique et à un allongement général du cycle cellulaire. Contrairement aux études précédentes, le retard mitotique des mESC Seh1-/- n'est pas lié à une délocalisation du Y-complexe ni de la kinase Aurora B au niveau des kinétochores.Pour comprendre si ce retard est dû à un rôle dans les NPC ou au sein du complexe GATOR2, j'ai ensuite conçu une stratégie visant spécifiquement à altérer l'interaction entre Seh1 et Nup85 (∆N-Nup85), son partenaire direct dans le Y-complexe. Dans les mutants ∆N-Nup85 Seh1 n'est plus détectable au niveau des kinétochores et est largement délocalisé du NPC. Cependant, les mESCs ∆N-Nup85 ne présentent aucun défaut de division cellulaire ni pendant la différenciation. D’autre part, j'ai constitutivement inactivé Mios, un autre membre du complexe GATOR2. Les cellules Mios-/- résultantes survivent pendant la différenciation et aucun retard de croissance cellulaire n'a été observé. En plus des défauts mentionnés précédemment, nous avons observé une diminution inattendue de la densité des NPC dans les cellules Seh1-/- et ∆N-Nup85, qui n'a cependant pas été observée dans le mutant Mios. Ceci indique une fonction de Seh1 dans la régulation de l'assemblage des NPC. Remarquablement, nous avons également observé une réduction de la taille dues noyaux des mutants Seh1 qui n'est pas observée dans les mutants ∆N Nup85, ce qui pourrait refléter un rôle de Seh1 dans le complexe GATOR2. Mes recherches ont élargi nos connaissances sur les différents rôles de Seh1 et ont commencé à aborder la localisation critique de cette protéine.The nuclear pore complexes (NPCs) are large macromolecular structures embedded in the nuclear envelope, composed of around 30 different proteins called nucleoporins or Nups. Besides their function in regulating nucleo-cytoplasmic transport, several Nups have been reported to play essential roles in other unrelated processes such as chromatin organization, gene regulation, cell cycle control and cell differentiation. The Nup107 complex (also termed Y-complex) is the major structural component of the NPC and is composed of 9 distinct Nups. The Y-complex has been shown to also localize at kinetochores, where it contributes to cell cycle progression. Moreover, one of the components of the Y-complex, Nup133, is required for embryonic stem cell differentiation and proper mouse development. More recent data further indicated a function of Nup133 in the assembly of the nuclear pore basket that may underlie some of its functions. The Y-complex has thus appeared as a key player in multiple biological processes. However, the specific roles of each of its subunits have just started to be investigated. Among the subunits of the Y-complex, Seh1 has been previously reported to play a role in chromosome alignment and segregation in cancer cell lines, by recruiting the Y-complex and several key mitotic regulators (including notably Aurora B) to kinetochores. Interestingly, Seh1 also belongs to the unrelated GATOR2 complex, an essential activator of the mTORC1 kinase, a master regulator of cell growth and proliferation. It is however unknown whether the mitotic functions of Seh1 rely on its localization at the pore, at kinetochores, or the GATOR2-complex. Due to the increasing number of studies implicating nuclear pore components in regulatory processes, Seh1 appears as a good candidate for the coordination of cell cycle and possibly differentiation. My data has revealed that Seh1-/- mESCs are viable in pluripotent state, but that cell survival is severely impaired upon differentiation. Pluripotent Seh1-/- mESCs also feature a cell growth rate delay when in the pluripotent state, correlated with a small (10 min) mitotic delay and and lenghthening of the interphase lenght as compared to WT mESCs. In contrast to previous studies performed in HeLa cells, we observed that both the Y-complex and Aurora B are still properly targeted to kinetochores in these cells. To understand if this delay is due to a role at NPCs or within the GATOR2 complex, I designed a strategy to specifically impair the interaction between Seh1 and Nup85 (∆N-Nup85), its direct partner within the Y-complex. In the resulting ∆N-Nup85 mutants, Seh1 is no longer detectable at kinetochores and is largely mislocalized from the NPC. In parallel, I constitutively inactivated Mios, another member of the GATOR2 complex. Surprisingly, the resulting ∆N-Nup85 and Mios-/- cells did progress through differentiation and no cell growth delay was observed. Interestingly, we observed an unexpected decrease in NPC density in the Seh1-/- and ∆N-Nup85 mESCs that was in contrast not observed in the Mios-/-cells. This pointed to a function of Seh1 in the context of the Y-complex in the regulation of NPC assembly.We also observed a reduction in nuclear size in the Seh1 mutants that is not shared by the ∆-Nup85mutants but that could be consistent with a role of Seh1 within the GATOR2 complex. My research has expanded our knowledge regarding the different roles of Seh1 and has started to tease apart the relative contributions of this protein in its different subcellular contexts

Integrity of the short arm of nuclear pore Y-complex is required for mouse embryonic stem cell growth and differentiation

Many cellular processes, ranging from cell division to differentiation, are controlled by nuclear pore complexes (NPCs). However, studying the contributions of individual NPC subunits to these processes in vertebrates has long been impeded by their complexity and the lack of efficient genetic tools. Here, we use genome editing in mouse embryonic stem cells (mESCs) to characterize the role of NPC structural components, focusing on the short arm of the Y-complex that comprises Nup85, Seh1 and Nup43. We show that Seh1 and Nup43, although dispensable in pluripotent mESCs, are required for their normal cell growth rates, their viability upon differentiation and for the maintenance of proper NPC density. mESCs with an N-terminally truncated Nup85 mutation (in which interaction with Seh1 is greatly impaired) feature a similar reduction of NPC density. However, their proliferation and differentiation are unaltered, indicating that it is the integrity of the Y-complex, rather than the number of NPCs, that is critical to ensure these processes

Androgen receptor gene status in plasma DNA associates with worse outcome on enzalutamide or abiraterone for castration-resistant prostate cancer: a multi-institution correlative biomarker study

BACKGROUND: There is an urgent need to identify biomarkers to guide personalized therapy in castration-resistant prostate cancer (CRPC). We aimed to clinically qualify androgen receptor (AR) gene status measurement in plasma DNA using multiplex droplet digital PCR (ddPCR) in pre- and post-chemotherapy CRPC. METHODS: We optimized ddPCR assays for AR copy number and mutations and retrospectively analyzed plasma DNA from patients recruited to one of the three biomarker protocols with prospectively collected clinical data. We evaluated associations between plasma AR and overall survival (OS) and progression-free survival (PFS) in 73 chemotherapy-naïve and 98 post-docetaxel CRPC patients treated with enzalutamide or abiraterone (Primary cohort) and 94 chemotherapy-naïve patients treated with enzalutamide (Secondary cohort; PREMIERE trial). RESULTS: In the primary cohort, AR gain was observed in 10 (14%) chemotherapy-naïve and 33 (34%) post-docetaxel patients and associated with worse OS [hazard ratio (HR), 3.98; 95% CI 1.74–9.10; P A (p.L702H) and 2632A>G (p.T878A)] were observed in eight (11%) post-docetaxel but no chemotherapy-naïve abiraterone-treated patients and were also associated with worse OS (HR 3.26; 95% CI 1.47–not reached; P = 0.004). There was no interaction between AR and docetaxel status (P = 0.83 for OS, P = 0.99 for PFS). In the PREMIERE trial, 11 patients (12%) with AR gain had worse PSA-PFS (sPFS) (HR 4.33; 95% CI 1.94–9.68; P < 0.001), radiographic-PFS (rPFS) (HR 8.06; 95% CI 3.26–19.93; P < 0.001) and OS (HR 11.08; 95% CI 2.16–56.95; P = 0.004). Plasma AR was an independent predictor of outcome on multivariable analyses in both cohorts. CONCLUSION: Plasma AR status assessment using ddPCR identifies CRPC with worse outcome to enzalutamide or abiraterone. Prospective evaluation of treatment decisions based on plasma AR is now required. CLINICAL TRIAL NUMBER: NCT02288936 (PREMIERE trial)